Crystalline forms of a macrolide, and uses therefor

ABSTRACT

New crystalline forms of macrolide compounds, and pharmaceutical compositions thereof, are described herein. In addition, processes for preparing the crystalline forms are described herein.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. provisional application61/316,063, filed 22 Mar. 2010, which is incorporated by referenceherein.

TECHNICAL FIELD

The invention described herein relates to a macrolide compound. Moreparticularly, it relates to new crystalline forms of the macrolidecompound, to processes for the preparation of the crystalline forms, andto pharmaceutical compositions containing the forms.

BACKGROUND AND SUMMARY OF THE INVENTION

It is appreciated that compounds considered as candidates for furtherdevelopment as pharmaceuticals advantageously possess desirablebiological properties, but also physical properties that adapt them foruse in the manufacture of pharmaceutical products. For example,compounds that form stable solids, including crystalline solids may bemore readily manufactured and formulated. It is further appreciated thatindividual physical forms of the compound that are stable andadditionally that may be prepared substantially free of other physicalforms may also be more readily manufactured and formulated. It is to beunderstood herein that different physical forms may have markedlydifferent physical properties, such as different solubilitycharacteristics, different bioavailabilities and/or biological exposure,different stability, and the like.

In US patent application publication number US 2006/0100164, there aredisclosed certain macrolide antibiotic compounds. The foregoingpublication, and each additional publication cited herein isincorporated herein by reference. One of these macrolides is afluoroketolide having Chemical Abstracts Registry Number 760981-83-7,which is also known as CEM-101 and solithromycin. The preparation of anamorphous form of CEM-101 is described therein. An alternativepreparation of CEM-101 is described in WO 2009/055557. CEM-101 has thefollowing chemical structure:

It has been discovered herein that CEM-101 can be isolated in a varietyof crystalline forms which provide illustrative embodiments of theinvention. CEM-101 can be isolated in crystalline form as a materialhaving a range of different physical properties, depending upon themethod of isolation. This is because CEM-101 can exist in more than onecrystalline form, i.e., it exhibits polymorphism. CEM-101 can beisolated in at least two crystalline forms, denoted herein as Form I andForm II, each of which is pure or substantially pure and/or free of orsubstantially free of the other form. Various mixtures of Form I andForm II can also be isolated. In addition, solids which are mixtures ofone ore more crystalline materials and also include amorphous solids canbe obtained.

BRIEF DESCRIPTION OF THE DRAWINGS

Illustrative examples of X-ray powder diffraction (XRPD) spectra foreach of Form I and Form II are given in FIG. 1 and FIG. 2, respectively,showing degrees 2θ on the X-axis and relative intensity on the Y-axis.

More detailed listings of the peaks for each of Form I and Form II areprovided below in Tables 1-4 in the Examples, in which peaks are denotedas % relative intensity (I/I₀×100). It is to be understood that in theX-ray powder diffraction spectra the exact values measured for °2θ (orthe corresponding d-spacings) may vary depending upon the particularsample analyzed and the particular analysis procedure used. A range ofvalues of at least ±0.1 °2θ, and in some cases at least ±0.2 °2θ, may betypical. Measurements on independently prepared samples on differentinstruments may lead to variability which is greater than +0.1 °2θand/or ±0.2 °2θ.

DETAILED DESCRIPTION

The new physical form, Form I of CEM-101 is a crystalline form which maybe described by its X-ray powder diffraction pattern. Peaks atapproximate positions of about ° 2θ=8.8, 10.5, 13.2 and 18.6 arerepresentative of this crystalline form. An example of an X-ray powderdiffraction spectrum for Form I is shown in FIG. 1. A more detailedanalysis of the peaks is described in Tables 1-2 below. Form I ofCEM-101 demonstrates minimal weight loss by thermogravimetric analysis(TGA). Water determination by Karl Fisher titration in typical lots is0.6 to 1.1%. Dynamic vapor sorption indicates that the material ispartially hygroscopic, but Form I solids are isolated upon desorption.Form I of CEM-101 melts at about 200° C. by DSC analysis.

As one embodiment there is described Form I of CEM-101 having an X-raypowder diffraction pattern with peaks at approximate positions of about2θ=8.8, 10.5, 13.2 and 18.6. As another embodiment there is described asolid form of CEM-101 comprising Form I of CEM-101 having an X-raypowder diffraction pattern with peaks at approximate positions of about° 2θ=8.8, 10.5, 13.2 and 18.6. As another embodiment there is describeda composition comprising CEM-101, where the majority of CEM-101 is FormI of CEM-101 having an X-ray powder diffraction pattern with peaks atapproximate positions of about ° 2θ=8.8, 10.5, 13.2 and 18.6. As anotherembodiment there is described CEM-101 in each of the foregoingembodiments having an X-ray powder diffraction pattern substantially thesame as that of FIG. 1.

The new physical form, Form I, is physically stable and can be preparedsubstantially free of other physical forms, as described below. In oneembodiment, there is described Form I of CEM-101 that is substantiallyfree of Form II. The relative amounts of forms may be determined by avariety of analytical techniques, for example but not limited to, by thepresence or absence of representative peaks, and/or by the relative peakheights and/or peak areas of appropriate discernable peaks in the XRPDspectrum. Form I of CEM-101, substantially free of other physical forms,may be characterized by an X-ray powder diffraction pattern with peaksat approximate positions of about °2θ=8.8, 10.5, 13.2 and 18.6. Peaks atabout °2θ=6.2, 19.7 and/or 21.9 are also observed for this crystallineform. In another embodiment, there is described Form I of CEM-101 ineach of the foregoing embodiments that is characterized by an X-raypowder diffraction pattern with peaks at approximate positions of about°2θ=8.8, 10.5, 13.2 and 18.6, and one or more additional peaks at about°2θ=6.2, 19.7 and/or 21.9. In another embodiment, there is describedForm I of CEM-101 in each of the foregoing embodiments that issubstantially free of Form II as determined by the X-ray powderdiffraction pattern, wherein one or more peaks at °2θ=5.6, 9.8, and/or11.7 are absent or nearly absent.

In another embodiment, pharmaceutical compositions that include CEM-101are described, where the composition comprises or consists essentiallyof Form I of CEM-101, as described in each of the foregoing embodiments.In another embodiment, pharmaceutical compositions that include CEM-101are described, where the composition comprises CEM-101, where theCEM-101 is at least about 60%, at least about 80%, at least about 90%,at least about 95%, at least about 98%, or at least about 99% Form I. Inanother embodiment, pharmaceutical compositions that include CEM-101 aredescribed, where the composition comprises CEM-101, where the CEM-101 issubstantially free of Form II. In another embodiment, pharmaceuticalcompositions that include Form I of CEM-101 are described, where therelative amount of Form II is less than about 40%, 20%, 10%, 5%, 2% or1%. In another embodiment, pharmaceutical compositions that includeCEM-101 are described, where the relative amount of 15 Form I and FormII is determined by the relative peak heights of specific peaks.Illustrative relative peak height ratios for the peaks at 2θ=6.2 and °2θ=5.6 are a ratio of about 5:1 or more, about 10:1 or more, or about20:1 or more. As another embodiment there is described Form I ofCEM-101, substantially free of other physical forms. As anotherembodiment there is described a pharmaceutical composition comprisingCEM-101 having an X-ray powder diffraction pattern substantially thesame as that of FIG. 1.

The new physical form, Form II of CEM-101 is a crystalline form whichmay be described by its X-ray powder diffraction pattern. Peaks atapproximate positions of about °2θ=5.6, 7.9, 9.3, 11.7, 12.9 and 16.7are representative of this crystalline form. An example of an X-raypowder diffraction spectrum for Form H is shown in FIG. 2. A moredetailed analysis of the peaks is described in Tables 3-4 below. Basedon a single crystal X-ray determination and dynamic vapor sorptionanalysis, Form II appears to be a non-hygroscopic anhydrous crystallineform that melts at about 225° C. as a single endothermic event by DSCanalysis. It is appreciated herein that non-hygroscopic solids, such asForm II described herein, may be advantageous in the preparation ofpharmaceutical compositions. Such advantages include improved handlingand stability properties in manufacturing, improved lot to lot qualitycontrol.

As one embodiment there is described Form II of CEM-101 having an X-raypowder diffraction pattern with peaks at approximate positions of about° 2θ=5.6, 7.9, 9.3, 11.7, 12.9 and 16.7. As another embodiment there isdescribed a solid form of CEM-101, comprising Form II of CEM-101 havingan X-ray powder diffraction pattern with peaks at approximate positionsof about °2θ=5.6, 7.9, 9.3, 11.7, 12.9 and 16.7. As another embodimentthere is described a composition comprising CEM-101, where the majorityof CEM-101 is Form II of CEM-101 having an X-ray powder diffractionpattern with peaks at approximate positions of about °2θ=5.6, 7.9, 9.3,11.7, 12.9 and 16.7. As another embodiment there is described CEM-101 ineach of the foregoing embodiments having an X-ray powder diffractionpattern substantially the same as that of FIG. 2.

The new physical form, Form II, is physically stable and can be preparedsubstantially free of other physical forms, as described below. In oneembodiment, there is described Form II of CEM-101 that is substantiallyfree of Form I. The relative amounts of forms may be determined by avariety of analytical techniques, for example but not limited to, by thepresence or absence of representative peaks, and/or by the relative peakheights and/or peak areas of appropriate discernable peaks in the XRPDspectrum. Form II of CEM-101, substantially free of other physicalforms, may be characterized by an X-ray powder diffraction pattern withpeaks at approximate positions of about °2θ=5.6, 7.9, 9.3, 11.7, 12.9and 16.7. In another embodiment, there is described Form II of CEM-101in each of the foregoing embodiments that is substantially free of FormI as determined by the X-ray powder diffraction pattern, wherein one ormore peaks at ° 2θ=6.2 and/or 8.8 are absent or nearly absent.

In another embodiment, pharmaceutical compositions that include CEM-101are described, where the composition comprises or consists essentiallyof Form II of CEM-101, as described in each of the foregoingembodiments. In another embodiment, pharmaceutical compositions thatinclude CEM-101 are described, where the composition comprises CEM-101,where the CEM-101 is at least about 60%, at least about 80%, at leastabout 90%, at least about 95%, at least about 98%, or at least about 99%Form II. In another embodiment, pharmaceutical compositions that includeForm II of CEM-101 are described, where the relative amount of Form I isless than about 40%, 20%, 10%, 5%, 2% or 1%. In another embodiment,pharmaceutical compositions that include CEM-101 are described, wherethe composition comprises CEM-101, where the CEM-101 is substantiallyfree of Form I. In another embodiment, pharmaceutical compositions thatinclude CEM-101 are described, where the relative amount of Form II andForm I is determined by the relative peak heights of specific peaks.Illustrative relative peak height ratios for the peaks at ° 2θ=5.6 and °2θ=6.2 are a ratio of about 5:1 or more, about 10:1 or more, or about20:1 or more. As another embodiment there is described Form II ofCEM-101, substantially free of other physical forms. Form II of CEM-101,substantially free of other physical forms, may be characterized by anX-ray powder diffraction pattern with peaks at approximate positions ofabout ° 2θ=5.6, 7.9, 9.3, 11.7, 12.9 and 16.7. As another embodimentthere is described a pharmaceutical composition comprising CEM-101having an X-ray powder diffraction pattern substantially the same asthat of FIG. 2.

As a further embodiment there is described a pharmaceutical compositioncomprising CEM-101 in any ratio of Form I and Form II. As a furtherembodiment there is described a pharmaceutical composition comprisingCEM-101 in any ratio of Form I and amorphous CEM-101. As a furtherembodiment there is described a pharmaceutical composition comprisingCEM-101 in any ratio of Form II and amorphous CEM-101. As a furtherembodiment there is described a pharmaceutical composition comprisingCEM-101 in any ratio of Form I and Form II and amorphous CEM-101.

Form I of CEM-101, including Form I substantially free of other physicalforms, may be prepared by recrystallization as described below in theExamples, for example by a recrystallization procedure as described inTable A., Experiments 18, 21-23 and 26. In the table, the ratios ofsolvent to CEM-101 are given in a volume:weight ratio (mL/mg or L/g)where “T” indicates “times” of volume. In general, Form I may beobtained by adding a solution of CEM-101 in a water miscible, polarorganic solvent, such as for example acetone, methanol or ethanol, towater to afford Form I of CEM-101.

According to one embodiment, described herein is a process for thepreparation of Form I of CEM-101. The process includes the step ofadding a solution of CEM-101 in a water miscible, polar organic solventto water, such as at a temperature below 50° C. In addition, the processmay include one or more of the following additional steps: heating thesolution of CEM-101, filtering the solution of CEM-101, reducing thevolume of the solution of CEM-101 by evaporation, stirring the waterduring the addition of the solution of CEM-101. In another embodiment,the prepared Form I of CEM-101 is substantially free of other physicalforms. In another embodiment, the prepared Form I of CEM-101 issubstantially free of Form II.

According to another embodiment, described herein is a process for thepreparation of Form I of CEM-101, which comprises one or more of thesteps of dissolving a source of CEM-101 in acetone, methanol or ethanol,or a combination thereof, optionally above ambient temperature,optionally filtering the solution, reducing the volume of the resultantsolution by evaporation, adding the solution to water at a temperaturebelow 50° C., optionally with stirring, and collecting the resultingcrystalline solid. A further embodiment is one wherein the above solventis acetone. A further embodiment is one wherein the above solvent isethanol.

For any of the above processes a further embodiment is one wherein thesolution is added to water at a temperature of about 10° C. to about 30°C. Another embodiment is one wherein the solution is added to waterdropwise. In another embodiment the prepared Form I of CEM-101 issubstantially free of other physical forms. In another embodiment theprepared Form I of CEM-101 is substantially free of Form II.

For any of the above processes for the preparation of Form I of CEM-101,any of the solvents acetone, methanol or ethanol, or a combinationthereof may be used. In another embodiment, the organic solution isslowly added to water at about 20-30° C. In another embodiment, thevolume:volume ratio of the organic solution to water is about 6 to about15. In another embodiment, the volume:volume ratio of the organicsolution to water is about 10 to about 13.

The above processes may be carried out with or without using seeds ofForm I of CEM-101.

Form II of CEM-101, including Form II substantially free of otherphysical forms, may be prepared as described below in the Examples, forexample by recrystallization as described in Table A., Experiments 1, 5,6, 8, 9, 11, 12, 14, 16, 19 and 24, or by a slurry procedure asillustrated in Table B. In general, Form II may be obtained by addingwater to a solution of CEM-101 in a water miscible, polar organicsolvent. In addition, Form II of CEM-101 may be obtained byrecrystallization from a number of organic solvents, with and withoutuse of an antisolvent, and with or without use of seeding, as shown inTable A. Alternatively, a mixture of Form I and Form II may be convertedinto Form II of CEM-101 by slurrying the mixture in, for example,2-propanol (isopropyl alcohol, IPA) at 60° C., or by slurrying themixture in 2-butanone (methyl ethyl ketone, MEK) under a variety ofconditions.

According to one embodiment, described herein is a process for thepreparation of Form II of CEM-101. The process includes the step ofadding water to a solution of CEM-101 in a water miscible, polar organicsolvent. In addition, the process may include one or more of thefollowing additional steps: filtering the solution of CEM-101, reducingthe volume of the solution of CEM-101 by evaporation, stirring thesolution of CEM-101 during addition of the water, and collecting theresulting crystalline solid. In another embodiment, the water miscible,polar organic solvent is protic. In another embodiment, the watermiscible, polar organic solvent is aprotic. A further embodiment is onewherein the water miscible, polar organic solvent is acetone,acetonitrile, 1,4-dioxane, methanol or ethanol, or a combinationthereof. A further embodiment is one wherein the solution of CEM-101 isabove ambient temperature, for example about 65 to about 80° C. or about65° C. A further embodiment is one wherein water at about ambienttemperature is added to the solution. Another embodiment is one whereinwater is added to the solution dropwise. In one embodiment of any of theabove processes for the preparation of Form II of CEM-101 thevolume:volume ratio of organic solvent to water is from about 1 to about10. In another embodiment the prepared Form II of CEM-101 issubstantially free of other physical forms.

The above processes may be carried out with or without using seeds ofForm II of CEM-101.

For the above procedures, when a source of CEM-101 is a solid, it may beCEM-101 in amorphous form, as Form I or Form II, as a furthercrystalline form, as a glass, or as a mixture of any of those forms. Itis to be understood that a solution of CEM-101 may also provide a sourceof CEM-101. As another embodiment, there is described a process ofpurifying CEM-101 comprising converting one or more forms or mixtures offorms of the CEM-101 into Form I or Form II substantially free of otherphysical forms. After such purification, it may be desired to convertCEM-101 into a different physical form for further use.

In another embodiment, described herein is a solid form of CEM-101prepared by a process that includes the step of adding a solution ofCEM-101 in a water miscible, polar organic solvent to water, such as ata temperature below 50° C. In addition, the process may include one ormore of the following additional steps: optionally with heating,optionally filtering the solution, optionally reducing the volume of theresultant solution by evaporation, optionally with stirring, andcollecting the resulting crystalline solid. In another embodiment, thewater is at a temperature of about 10° C. to about 30° C. In anotherembodiment, the solvent is acetone, methanol or ethanol, or acombination thereof. In another embodiment, the organic solution isslowly added to water at about 20-30° C. In another embodiment, thevolume:volume ratio of the organic solution to water is about 6 to about15. In another embodiment, the volume:volume ratio of the organicsolution to water is about 10 to about 13. In variations of the above,the process may include using seeds of Form I of CEM-101.

In another embodiment, described herein is a solid form of CEM-101prepared by a process that includes the step of adding water to asolution of CEM-101 in a water miscible, polar organic solvent. Inaddition, the process may include one or more of the followingadditional steps: optionally filtering the solution, optionally reducingthe volume of the resultant solution by evaporation, optionally withstirring, and collecting the resulting crystalline solid. In anotherembodiment, the water miscible, polar organic solvent is protic. Inanother embodiment, the water miscible, polar organic solvent isaprotic. A further embodiment is one wherein the water miscible, polarorganic solvent is acetone, acetonitrile, 1,4-dioxane, methanol orethanol, or a combination thereof. A further embodiment is one whereinthe solution of the water miscible, polar organic solvent is aboveambient temperature, for example about 65 to about 80° C. or about 65°C. A further embodiment is one wherein water at about ambienttemperature is added to the solution. Another embodiment is one whereinwater is added to the solution dropwise. In another embodiment, thevolume:volume ratio of organic solvent to water is from about 1 to about10. In variations of the above, the process may include using seeds ofForm II of CEM-101.

As another embodiment, there is described a pharmaceutical compositioncomprising CEM-101 in crystalline form as described in any of thedescriptions herein and further comprising at least one pharmaceuticallyacceptable carrier or excipient.

Illustratively, compositions may include one or more carriers, diluents,and/or excipients. The compounds described herein, or compositionscontaining them, may be formulated in a therapeutically effective amountin any conventional dosage forms appropriate for the methods describedherein, and include one or more carriers, diluents, and/or excipientstherefor. Such formulation compositions may be administered by a widevariety of conventional routes for the methods described herein, and ina wide variety of dosage formats, utilizing known procedures. Capsulesand tablets are embodiments commonly used for oral administration ofantibiotics. See generally, Remington: The Science and Practice ofPharmacy, (21^(st) ed., 2005), as well as an illustrative formulationcomposition in the Examples.

As another embodiment, there is described a method of treatment of abacterial infection, a protozoal infection, or a disorder related to abacterial infection or protozoal infection comprising the step ofadministering to a subject in need thereof a therapeutically effectiveamount of CEM-101 in crystalline form as described herein, or apharmaceutical composition thereof further comprising at least onepharmaceutically acceptable carrier or excipient. Illustrative dosingschedules include the daily administration of a composition, in a singleor divided format, comprising about 1,200 mg, about 1,000 mg, about 800mg, about 400 mg, about 200 mg, or about 100 mg.

As another embodiment, there is described a use of CEM-101 incrystalline form as described herein for the treatment of a bacterialinfection, a protozoal infection, or a disorder related to a bacterialinfection or protozoal infection.

As another embodiment, there is described a use of CEM-101 incrystalline form as described herein for the manufacture of a medicamentfor the treatment of a bacterial infection, a protozoal infection, or adisorder related to a bacterial infection or protozoal infection.

As a further embodiment, method or use described above is one whereinthe subject is a mammal, a fish, a bird or a reptile. As anotherembodiment, there is described a method or use wherein the subject is amammal. As another embodiment, there is described a method or usewherein the subject is a human.

The term “therapeutically effective amount” as used herein, refers tothat amount of active compound or pharmaceutical agent that elicits thebiological or medicinal response in a tissue system, animal or humanthat is being sought by a researcher, veterinarian, medical doctor orother clinician, which includes alleviation of the symptoms of thedisease or disorder being treated. In one aspect, the therapeuticallyeffective amount is that which may treat or alleviate the disease orsymptoms of the disease at a reasonable benefit/risk ratio applicable toany medical treatment. However, it is to be understood that the totaldaily usage of the compounds and compositions described herein may bedecided by the attending physician within the scope of sound medicaljudgment. The specific therapeutically-effective dose level for anyparticular patient will depend upon a variety of factors, including thedisorder being treated and the severity of the disorder, activity of thespecific compound employed; the specific composition employed; the age,body weight, general health, gender and diet of the patient: the time ofadministration, route of administration, and rate of excretion of thespecific compound employed; the duration of the treatment; drugs used incombination or coincidentally with the specific compound employed; andlike factors well known to the researcher, veterinarian, medical doctoror other clinician of ordinary skill.

As shown in the interconversion of slurries experiment summarized belowin the Examples in Table B, slurries of Forms I and II are prepared in2-propanol (IPA) and 4-butanone (MEK) at ambient, sub-ambient, andelevated temperatures. In 2-propanol, conversion of Form I to Form Iappears to be solubility driven at ambient and sub-ambient temperatures,with little apparent change observed in the ratio of Forms I and II byXRPD analysis over 3 days. Extending the interconversion period to 8days at room temperature left only a trace of Form I remaining. In2-propanol, at elevated temperature, only Form II is obtained.Experiments based on 2-butanone did not have the apparent solubilitylimitation and gave Form II as the only observed form at sub-ambient,ambient and elevated temperatures. Based on these experiments and theDSC results, but without being bound by theory, it appears that Form IImay be more stable than Form I throughout the temperature range. Inaddition, but without being bound by theory, the DSC results suggestthat Form I and II are monotropically related.

As shown below in the Examples in Table C-1, Form I exhibited greateraqueous solubility than Form II over a pH range from 9.2 to 1.2.

When the forms of CEM-101 are dosed in a vehicle in which the compoundis well solubilized, there is no observed difference in measuredpharmacokinetic parameters, including Cmax, Tmax, and AUC, among Form Iand Form II. When the forms of CEM-101 are dosed in a vehicle in whichthe compound is present as a solid, there is no statistical differencein measured pharmacokinetic parameters, including Cmax, Tmax, and AUC,among Form I and Form II.

In another embodiment, crystalline forms of CEM-101, and mixturesthereof, are described herein that show improved solubility compared toamorphous forms of CEM-101

Examples X-Ray Powder Diffraction (XRPD) Peak Positions.

XRPD patterns were collected using an Inel XRG-3000 diffractometerequipped with a curved position sensitive detector with a 20 range of120°. An incident beam of Cu Kα radiation (40 kV, 30 mA) was used tocollect data in real time at a resolution of 0.03 °2θ. Prior to theanalysis, a silicon standard (NIST SRM 640c) was analyzed to verify theSi 111 peak position. Samples were prepared for analysis by packing theminto thin-walled glass capillaries. Each capillary was mounted onto agoniometer head and rotated during data acquisition. The monochromatorslit was set at 5 mm by 160 μm.

Alternatively, XRPD patterns were collected using a PANalytical X'PertPro diffractometer. An incident beam of Cu Kα radiation was producedusing an Optix long, fine-focus source. An elliptically gradedmultilayer mirror was used to focus the Cu Kα X-rays of the sourcethrough the specimen and onto the detector. Data were collected andanalyzed using X'Pert Pro Data Collector software (v. 2.2b). Prior tothe analysis, a silicon specimen (NIST SRM 640c) was analyzed to verifythe Si 111 peak position. The specimen was sandwiched between 3 μm thickfilms, analyzed in transmission geometry, and rotated to optimizeorientation statistics. A beam-stop and a helium atmosphere were used tominimize the background generated by air scattering. Soller slits wereused for the incident and diffracted beams to minimize axial divergence.Diffraction patterns were collected using a scanning position-sensitivedetector (X'Celerator) located 240 mm from the specimen.

The data presented herein contain X-ray diffraction patterns and tableswith peak lists. It is to be understood that the range of data collectedmay be instrument dependent. Under typical circumstances, peaks withinthe range of up to about 30 °2θ are selected. Although peaks may belabeled on diffraction patterns and listed in tables, it is to beunderstood that different rounding algorithms may be used to round eachpeak to the nearest 0.1 or 0.01 °2θ, depending upon the instrument usedto collect the data and/or the inherent peak resolution. Should there bea difference, the peak positions listed in the tables should be used.The location of the peaks along the x-axis (°2θ) in both the figures andthe tables may be automatically determined using appropriate software,which is typically resident on the instrument used and rounded to one ortwo significant figures after the decimal point based upon the abovecriteria. Peak position variabilities are given herein to within ±0.1°2θ based upon recommendations outlined in the USP discussion ofvariability in x-ray powder diffraction (United States Pharmacopeia, USP32, NF 27, Vol. 1, pg. 392, 2009). The accuracy and precision associatedwith any particular measurement reported herein has not been determined.Measurements on independently prepared samples on different instrumentsmay lead to variability which is greater than ±0.1 °2θ. For d-spacelistings, the wavelength used to calculate d-spacings was 1.541874 Å, aweighted average of the Cu—K_(α1) and Cu—K_(α2) wavelengths (Phys. Rev.A56(6) 4554-4568 (1997)). Variability associated with d-spacingestimates was calculated from the USP recommendation, at each d-spacing,and provided in the respective data tables.

When multiple diffraction patterns are available, then assessments ofparticle statistics (PS) and/or preferred orientation (PO) are possible.Reproducibility among XRPD patterns from multiple samples analyzed on asingle diffractometer indicates that the particle statistics areadequate. Consistency of relative intensity among XRPD patterns frommultiple diffractometers indicates good orientation statistics.Alternatively, the observed XRPD pattern may be compared with acalculated XRPD pattern based upon a single crystal structure, ifavailable. Two-dimensional scattering patterns using area detectors canalso be used to evaluate PS/PO. If the effects of both PS and PO aredetermined to be negligible, then the XRPD pattern is representative ofthe powder average intensity for the sample and prominent peaks may beidentified as “Representative Peaks”. In general, it is appreciated thatthe more data collected to determine Representative Peaks, the morestatistically significant is the classification of those peaks.

CEM-101 Form I.

One PANalytical pattern and one Inel pattern were analyzed for thismaterial, and therefore at least partially optimized orientation andparticle statistic effects could be assessed through comparison withmultiple patterns. The peak positions and intensities are consistentbetween patterns, indicating adequate particle and orientationstatistics. Observed Peaks are shown in FIG. 1 and Table 1, andRepresentative Peaks are listed in Table 2.

TABLE 1 Observed peaks for CEM-101 Form I. °2θ d-space (Å) Intensity (%) 6.19 ± 0.10 14.268 ± 0.234  13  8.47 ± 0.10 10.443 ± 0.125  30  8.80 ±0.10 10.047 ± 0.115  68  9.34 ± 0.10 9.473 ± 0.102 10 10.52 ± 0.10 8.407± 0.080 100 10.97 ± 0.10 8.062 ± 0.074 46 12.04 ± 0.10 7.349 ± 0.061 2512.41 ± 0.10 7.132 ± 0.058 43 13.20 ± 0.10 6.709 ± 0.051 88 13.68 ± 0.106.472 ± 0.047 41 14.52 ± 0.10 6.102 ± 0.042 22 14.85 ± 0.10 5.965 ±0.040 22 17.02 ± 0.10 5.209 ± 0.031 45 18.03 ± 0.10 4.921 ± 0.027 4618.61 ± 0.10 4.768 ± 0.026 71 19.48 ± 0.10 4.557 ± 0.023 48 19.73 ± 0.104.500 ± 0.023 56 20.68 ± 0.10 4.294 ± 0.021 38 21.17 ± 0.10 4.197 ±0.020 39 21.89 ± 0.10 4.061 ± 0.018 53 23.41 ± 0.10 3.801 ± 0.016 2524.44 ± 0.10 3.642 ± 0.015 11 24.91 ± 0.10 3.574 ± 0.014 37 25.61 ± 0.103.478 ± 0.013 18 26.35 ± 0.10 3.383 ± 0.013 11 26.80 ± 0.10 3.327 ±0.012 24 29.04 ± 0.10 3.075 ± 0.010 18 29.96 ± 0.10 2.983 ± 0.010 10

TABLE 2 Representative peaks for CEM-101 Form I. °2θ d-space (Å)Intensity (%)  8.80 ± 0.10 10.047 ± 0.115  68 10.52 ± 0.10 8.407 ± 0.080100 13.20 ± 0.10 6.709 ± 0.051 88 18.61 ± 0.10 4.768 ± 0.026 71

CEM-101 Form II.

One Inel pattern was analyzed for this material, and preferredorientation and particle statistic effects could be assessed throughcomparison with the simulated pattern from single crystal data (notshown). The peak positions and intensities are consistent betweenpatterns, indicating adequate particle and orientation statistics.Observed peaks are shown in FIG. 2 and Table 3, and representative peaksare listed in Table 4.

TABLE 3 Observed peaks for CEM-101 Form II. °2θ d-space (Å) Intensity(%)  5.60 ± 0.10 15.772 ± 0.286  53  7.85 ± 0.10 11.260 ± 0.145  54 9.30 ± 0.10 9.505 ± 0.103 69  9.82 ± 0.10 9.004 ± 0.092 36 10.65 ± 0.108.304 ± 0.078 36 11.24 ± 0.10 7.871 ± 0.070 45 11.66 ± 0.10 7.592 ±0.065 75 11.97 ± 0.10 7.395 ± 0.062 38 12.87 ± 0.10 6.880 ± 0.054 10013.28 ± 0.10 6.666 ± 0.050 33 13.77 ± 0.10 6.432 ± 0.047 32 14.15 ± 0.106.260 ± 0.044 36 14.63 ± 0.10 6.054 ± 0.041 35 15.12 ± 0.10 5.861 ±0.039 48 16.71 ± 0.10 5.306 ± 0.032 78 17.23 ± 0.10 5.147 ± 0.030 3317.50 ± 0.10 5.067 ± 0.029 36 18.37 ± 0.10 4.830 ± 0.026 38 18.58 ± 0.104.776 ± 0.026 43 18.78 ± 0.10 4.724 ± 0.025 40 19.34 ± 0.10 4.590 ±0.024 37 19.68 ± 0.10 4.510 ± 0.023 46 20.62 ± 0.10 4.308 ± 0.021 3820.97 ± 0.10 4.237 ± 0.020 43 21.17 ± 0.10 4.196 ± 0.020 47 21.38 ± 0.104.156 ± 0.019 35 21.83 ± 0.10 4.071 ± 0.019 34 22.11 ± 0.10 4.021 ±0.018 34 22.59 ± 0.10 3.936 ± 0.017 27 23.32 ± 0.10 3.815 ± 0.016 3223.49 ± 0.10 3.787 ± 0.016 32 24.15 ± 0.10 3.685 ± 0.015 28 24.43 ± 0.103.644 ± 0.015 32 24.77 ± 0.10 3.594 ± 0.014 30 25.61 ± 0.10 3.479 ±0.013 30 25.78 ± 0.10 3.456 ± 0.013 29 26.23 ± 0.10 3.398 ± 0.013 2726.58 ± 0.10 3.354 ± 0.012 25 27.23 ± 0.10 3.275 ± 0.012 26 27.55 ± 0.103.238 ± 0.012 26 27.86 ± 0.10 3.203 ± 0.011 31 28.52 ± 0.10 3.130 ±0.011 24 29.49 ± 0.10 3.029 ± 0.010 26

TABLE 4 Representative peaks for CEM-101 Form II. °2θ d-space (Å)Intensity (%)  5.60 ± 0.10 15.772 ± 0.286  53  7.85 ± 0.10 11.260 ±0.145  54  9.30 ± 0.10 9.505 ± 0.103 69 11.66 ± 0.10 7.592 ± 0.065 7512.87 ± 0.10 6.880 ± 0.054 100 16.71 ± 0.10 5.306 ± 0.032 78

Recrystallization Studies

Recrystallization conditions that yield Form I and Form II, as well asconditions which afford amorphous material or mixtures of Form I andForm II, as well as the outputs (recoveries) from the conditions areshown in the following Table A. Recrystallization Studies. The polymorphform was determined by analysis of the X-ray powder diffraction patternfor each isolated product. In the table, ‘T’ represents the ratio ofsolvent to the mass of solid in mL/mg (or L/g); DCM meansdichloromethane; DMF means dimethylformamide; and IPE meansisopropylether.

TABLE A Recrystallization Study Results CEM- CEM-101 101 PolymorphExperiment Input Solvent Procedure Output Form 1  5 g Acetone Water(10T) Dissolved solid in acetone 4.5 g Form II   (5T) and added waterdropwise   at 30° C. 2  5 g DCM (5T) Dissolved in DM and 4.4 g Amorphous  freeze dried 3  5 g Acetone Water (50T) Dissolved solid in acetone 4.4g Form I +   (5T) and added to water Form II   dropwise at 30° C. 4  2 gDMF(2T) Water (50T) Dissolved solid in DMF 1.8 g Form I   and added towater   dropwise at 28° C. 5  3 g Methanol Water (18T) Dissolved solidin 2.5 g Form II   (18T) methanol and added water   dropwise at 28° C. 6 3 g Ethanol Water (10T) Dissolved solid in ethanol 2.2 g Form II  (10T) and added water dropwise   at 28° C. 7  3 g Methanol (18T)Dissolved in methanol and 2.7 g Form II +   freeze dried Form I 8  3 gDCM (3T) Cyclohexane Dissolved solid in DCM 2.3 g Form II   (20T) andadded to cyclohexane   in one portion at 28° C. 9  3 g Ethanol IPE (33T)Dissolved solid in ethanol 1.2 g Form II   (10T) and added to IPE in one  portion at 28° C. 10  2 g Acetonitrile (4T) Dissolved solid in 0.6 gForm I +   acetonitrile and cooled to Form II   30° C. 11  2 g1,4-Dioxane Water (5T) Dissolved solid in 1,4- 2.1 Form II   (5T)Dioxane at 70° C. and   added water at 0° C. to   5° C. 12  2 gAcetonitrile Water (4T) Dissolved solid in 1.6 g Form II   (4T)acetonitrile at 80° C. and   added water at 55° C. 13  1 g Acetonitrile(4T) Dissolved solid in 0.5 g Form II +   acetonitrile at 80° C. andForm I   seeded with 10 mg of   Form I 14  1 g Acetonitrile (4T)Dissolved solid in 0.4 g Form II acetonitrile at 80° C. and seeded with100 mg of Form I 15 12 g DCM (3T) Dissolved in DCM and 9.3 g Amorphousfreeze dried 16  1 g Ethanol Water (5T) Dissolved solid (Form I) 0.8 gForm II   (15T) in ethanol (15 mL) in   65° C., reduced volume to 5   mLand added water (5   mL) dropwise at 28° C. 17  2 g DMF(2T) Water (50T)Dissolved solid (Form II) 1.9 g Form I +   in DMF (4 mL) and added FormII   to water (100 mL)   dropwise at 28° C. 18  5 g Acetone Water (50T)Dissolved solid (Form II) 4.9 g Form I   (7T) in acetone (35 mL) in  65° C., reduced volume to 20   mL and added to water   (100 mL) dropwiseat   28° C.. The wet solid was   suspended in water and   stirred underheating at   65° C., cooled and filtered. 19  2 g Water Dissolved solid(Form II) 1.8 g Form II   in water (100 mL) and   stirred under heatingat   60° C., cooled and filtered. 20  5 g Ethanol Water (5T) Dissolvedsolid (Form II) 4.8 g Unknown   (15T) in ethanol (75 mL) at   65° C.,reduced volume to 35   mL and added water   dropwise at 60° C.   Ethanolwas removed by   distillation after formation   of the solid, stirringunder   heating, cooled and   filtered. 21  5 g Acetone Water (60T)Dissolved solid (Form 1 + 4.5 g Form I   (6T) II) in acetone (30 mL) at  65° C., filtered, reduced   volume to 20 mL and   added to water (300mL)   dropwise at 27° C., stirred   and filtered. 22  5 g Acetone Water(60T) Dissolved solid (Form I + 4.5 g Form I   (8T) II) in acetone (40mL) at   65° C., filtered, reduced   volume to 30 mL and   added towater (450 mL)   dropwise at 27° C., stirred   and filtered. 23  2 gEthanol Water (60T) Dissolved solid (Form I + 1.7 g Form I   (15T) II)in ethanol (30 mL) at   65° C., filtered, reduced   volume to 16 mL and  added to water (120 mL)   dropwise at 27° C., stirred   and filtered.24  5 g Acetone Water (60T) Dissolved solid (Form I + 4.4 g Form II  (6T) II) in acetone (30 mL) at   65° C., filtered, reduced   volume to20 mL and   water (300 mL) was added   dropwise at 27° C., stirred   andfiltered. 25  5 g Acetone Water (50T) Dissolved solid (Form I + 4.5 gForm I +   (7T) II) in acetone (35 mL) at little of   65° C., filtered,reduced Form II   volume to 25 mL and   added to water (250 mL)  dropwise at 27° C., stirred   and filtered. 26  5 g Ethanol Water (50T)Dissolved solid (Form I + 4.4 g Form I (15T) II) in ethanol (75 mL) at65° C., filtered, reduced volume to 40 mL and added to water (250 mL)dropwise at 27° C., stirred and filtered.

Hot Stage Microscopy and DSC

Form I of CEM-101 exhibits a melt onset at about 180° C. and final meltat about 200° C. by hot stage microscopy and exhibits endothermic eventsat 170 and 197-198° C. by differential scanning calorimetry (DSC). FormII of CEM-101 exhibits a melt onset at about 215° C. by hot stagemicroscopy and a DSC peak at about 225° C., as a single endothermicevent. Mixtures of varying amounts of Form I and Form II have exhibitedendothermic events at 194-199 and at 219-225° C. by DSC, depending uponthe ratio of the forms. It is to be understood that hot stage microscopyand/or DSC may be used to determine the presence or absence of certainforms in the compounds and compositions described herein.

Interconversion Studies.

Interconversion studies by slurrying of Form I and Form H are shown inthe following Table B in which the starting solid was a mixture of FormI and Form II; IPA is isopropyl alcohol (isopropanol) and MEK is methylethyl ketone (2-butanone).

Interconversion Slurries for CEM-101 Form A and B Solids XRPD SolventConditions^(a) Result^(b) IPA 60° C. 3 days Form II RT 3 days Form I +II RT 5 days ^(c) (8 days total) Form II + trace Form I 2-8° C. 3 daysForm I + II 2-8° C. 5 days ^(d) Form I + II (8 days total) MEK 6° C. 3days IS^(b) 60° C. 3 days ^(c) (6 days total) Form II RT 3 days Form II2-8° C. 3 days Form II ^(a)Reported times and temperatures areapproximate. RT = ambient temperature. ^(b)XRPD = X-ray powderdiffraction, IS = insufficient solids for analysis. ^(c.) Continuationof previous slurry with additional solids. ^(d.) Continuation ofprevious slurry.

Aqueous Solubility.

Using a linear calibration curve obtained by plotting the concentrationof CEM-101 vs. LC/MS-MS peak areas, the aqueous solubility of forms ofCEM-101 are obtained as follows: Test compounds (dry powder) are addedto 0.9% saline (initial pH 6.03) until precipitation, and the mixture isincubated for 6 h with shaking. After the incubation period, the samplesare centrifuged twice and solubility is estimated using the linearitystandard curve for the compound: Form I showed a solubility of 1411μg/mL.

Using a linear calibration curve obtained by plotting the concentrationof CEM-101 vs. LC/MS-MS peak areas, the aqueous solubility of forms ofCEM-101 was obtained as follows: Test compounds (dry powder) were addedto water with the indicated pH until precipitation, and the mixture wasincubated for 6 h with shaking. After the incubation period, the sampleswere centrifuged twice and solubility was estimated using the linearitystandard curve for the compound: Media: Water at pH: pH 9.2, 7.4, 4 and1.2 (adjusted using 0.1N NaOH and 0.1 N HCl); Incubation: 6 h at 26° C.with shaking; Test concentrations: Dry powder added till saturation;Detection: LC/MS-MS. The results are shown in Table C-1.

TABLE C-1 Aqueous Solubility Study Form II Form I pH (μg/mL) (μg/mL) 9.2331 1751 7.4 305 2224 4 361 4656 1.2 3638 4808

Intrinsic Dissolution Comparison.

Mean intrinsic dissolution rates for exemplary batches of the two formsare shown in Table cC-2 for 3 batches of Form I (SD and % CV for onebatch shown) and for Form II.

TABLE C-2 Mean Intrinsic Dissolution Studies. Mean intrinsic dissolutionrate (mg/min/cm²) Form I Form II Dissolution Medium Batch A Batch BBatch C SD* % CV* SD % CV 0.1N Hydrochloric Acid 9.4 10.6 7.9 35.8 9.88.8 38.0 9.4 Acetate buffer, pH 4.5 2.1 2.8 2.7 6.43 5.2 2.7 6.4 10.3Phosphate buffer, pH 6.8 0.004 0.03 0.02 0.53 53.0 0.01 0.5 51.1Phosphate buffer, pH 7.6 0.02 0.001 ND NA NA ND NA NA Water 2.0 0.7 1.64.91 9.6 ND NA NA *SD and CV % for Batch C only. ND, not determined. NA,not applicable.

Pharmacokinetic Evaluation of CEM-101 in Balb/c Mice.

The pharmacokinetics of lots of CEM-101 characterized as Form I and FormII were evaluated in Balb/c mice receiving a singe dose of 20 or 100mg/kg body weight (b.w.) via oral gavage. The doses were prepared as 2.0mg/mL or 10.0 mg/mL in a vehicle of 0.5% (w/v) carboxymethyl cellulosein water and a dose volume of 10 mL/kg b.w. Blood samples were collectedat various time points during the next 6 h post dose. The results areshown as follows:

TABLE D Plasma concentrations of CEM-101 in μg/mL vs. time in femaleBalb/c mouse at 20 mg/kg b.w. CEM-101 Form I and Form II Polymorphs Oralpharmacokinetics studies in female Balb/c mouse (20 mg/kg b.w.) Plasmaconcentrations (μg/mL); Female Balb/c mouse (20 mg/kg b.w.) CEM-101FormII CEM-101 Form I Time (h) Mean Stdv Mean Stdv 0.00 0.0 ± 0.0 0.0 ± 0.00.25 6.2 ± 1.6 8.2 ± 2.4 0.50 8.8 ± 0.8 13.6 ± 2.0 1.00 7.3 ± 0.9 12.0 ±1.7 2.00 6.1 ± 1.0 11.3 ± 1.4 4.00 5.5 ± 1.7 7.7 ± 0.7 6.00 1.3 ± 0.54.2 ± 0.4

TABLE E Plasma concentrations of CEM-101 in μg/mL vs. time in maleBalb/c mouse at 20 mg/kg b.w. CEM-101 Form I and Form II Polymorphs Oralpharmacokinetics studies in male Balb/c mouse (20 mg/kg b.w.) Plasmaconcentrations (μg/mL); Male Balb/c mouse (20 mg/kg b.w.) CEM-101 FormII CEM-101 Form I Time (h) Mean Stdv Mean Stdv 0.00 0.0 ± 0.0 0.0 ± 0.00.25 4.1 ± 0.8 5.3 ± 1.8 0.50 6.7 ± 0.6 7.7 ± 1.6 1.00 5.9 ± 0.8 9.5 ±0.9 2.00 5.6 ± 0.4 6.4 ± 0.8 4.00 4.9 ± 1.1 5.9 ± 0.6 6.00 2.9 ± 0.6 2.6± 0.8

TABLE F Plasma concentrations of CEM-101 in μg/mL vs. time in femaleBalb/c mouse at 100 mg/kg b.w. CEM-101 Form I and Form II PolymorphsOral pharmacokinetics studies in female Balb/c mouse (100 mg/kg b.w.)Plasma concentrations (μg/mL); Female Balb/c mouse (100 mg/kg b.w.)CEM-101 Form II CEM-101 Form I Time (h) Mean Stdv Mean Stdv 0.00 0.0 ±0.0 0.0 ± 0.0 0.25 7.3 ± 0.9 11.9 ± 1.7 0.50 10.5 ± 1.4 11.6 ± 1.6 1.009.6 ± 2.2 14.7 ± 2.3 2.00 9.5 ± 2.2 14.1 ± 2.9 4.00 10.2 ± 1.0 16.0 ±2.3 6.00 7.8 ± 1.1 19.5 ± 2.3

TABLE G Plasma concentrations of CEM-101 in μg/mL vs. time in maleBalb/c mouse at 100 mg/kg b.w. CEM-101 Form I and Form II PolymorphsOral pharmacokinetics studies in female Balb/c mouse (100 mg/kg b.w.)Plasma concentrations (μg/mL); Male Balb/c mouse (100 mg/kg b.w.)CEM-101 Form II CEM-101 Form I Time (h) Mean Stdv Mean Stdv 0.00 0.0 ±0.0 0.0 ± 0.0 0.25 6.8 ± 0.7 11.3 ± 2.5 0.50 6.9 ± 1.1 10.7 ± 3.0 1.008.7 ± 2:7 13.1 ± 3.2 2.00 9.3 ± 1.2 13.4 ± 3.1 4.00 10.6 ± 1.4 17.4 ±2.9 6.00 7.2 ± 0.8 12.0 ± 2.0

TABLE H Mean Plasma PK Parameters at 20 mg/kg. CEM-101 Form I and FormII Polymorphs Oral pharmacokinetics studies in Balb/c mouse Mean PlasmaPK Parameters Male-20 mg/kg Female-20 mg/kg CEM-101 CEM-101 CEM-101CEM-101 Parameters Form II Form I Form II Form I Route of Oral Oral OralOral administration Dose (mg/kg) 20 20 20 20 Cmax (μg/mL) 6.73 9.5 8.813.6 Tmax (h) 0.5 1 0.5 0.5 AUClast (h*μg/mL) 29 35 31.6 53 (0 to 6 h)AUCinf (h*μg/mL) 49 46 36 70 AUCextrap (%) 42 24 12 24

TABLE I Mean Plasma PK Parameters at 100 mg/kg. CEM-101 Form I and FormII Polymorphs Oral pharmacokinetics studies in Balb/c mouse Mean PlasmaPK Parameters Male-100 mg/kg Female-100 mg/kg CEM-101 CEM-101 CEM-101CEM-101 Parameters Form II Form I Form II Form I Route of administrationOral Oral Oral Oral Dose (mg/kg) 100 100 100 100 Cmax (μg/mL) 10.6 17.410.5 19.5 Tmax (h) 4 4 4 6 AUClast (h*μg/mL) (0 66 83.5 55 91 to 6 h)AUCinf (h*μg/mL) 99 NC NC NC AUCextrap (%) 33 NC NC NC

Pharmacokinetic (PK) Comparison.

Results of a further oral pharmacokinetic study of Form I and Form II ingroups of 5 female Balb/c mice receiving a singe dose at 5, 10 and 20mg/kg is are shown in Table J.

TABLE J Mean Plasma PK Parameters Dosing 5 mg/kg 10 mg/kg 20 mg/kg AUCAUC AUC Form c t½ Tmax Cmax 0-24 t½ Tmax Cmax 0-24 t½ Tmax Cmax 0-24 I N1 5 5 5 4 5 5 5 4 5 5 5 Mean 1.32 0.600 1.43 3.30 3.60 2.40 4.68 17.72.10 1.60 6.64 26.2 SD NC 0.224 0.249 0.551 1.65 0.894 1.31 3.25 0.5090.548 1.84 4.84 Min 1.32 0.500 1.02 2.78 2.44 2.00 3.21 14.4 1.73 1.004.66 19.5 Median 1.32 0.500 1.54 3.16 2.95 2.00 4.09 17.2 1.91 2.00 5.9126.7 Max 1.32 1.00 1.65 4.24 6.05 4.00 6.11 21.1 2.85 2.00 9.34 33.0 CV% NC 37.3 17.4 16.7 46.0 37.3 27.9 18.3 24.2 34.2 27.7 18.5 II N 5 5 5 53 5 5 5 2 5 5 5 Mean 1.17 1.20 1.85 4.69 1.42 1.25 2.63 7.58 2.46 1.105.36 20.1 SD 0.297 0.417 0.249 0.819 0.263 0.750 1.22 2.27 0.0517 0.5480.837 3.33 Min 0.935 1.00 1.55 4.21 1.12 0.250 1.50 4.62 2.43 0.500 3.8816.3 Median 1.12 1.00 1.80 4.31 1.53 1.00 2.42 7.59 2.46 1.00 5.66 21.7Max 1.67 2.00 2.12 6.14 1.61 2.00 4.69 10.0 2.50 2.00 5.96 23.4 CV %25.3 37.3 13.5 17.5 18.5 60.0 46.3 30.0 2.10 49.8 15.6 16.6

Examples of Pharmaceutical Compositions Using CEM-101 Form I.

The following formulation is used to provide Size 0 hard gelatincapsules containing 200 mg of CEM-101 per capsule.

Material % w/w Qty/Cap (mg) CEM-101 61.54 200.00 Avicel PH101 30.96100.63 Plasdone K29/32 2.00 6.50 Ac-Di-Sol 4.00 13.00 Sodium LaurylSulfate, NF 1.00 3.25 Magnesium Stearate, NF 0.50 1.63 (Vegetable Grade)Purified Water,* USP 64.3 — Total 100.00 325.00 *Estimated amount ofpurified water. Removed during processing; not in final formula.

The following formulation is used to provide tablets containing 200 mgper tablet, containing 54% drug load and a target weight of 370mg/tablet. The drug product is manufactured by wet granulation and thetablets are compressed at 6 kp.

Material % w/w Qty/Tablet(Ing) CEM-101* (Intra-granular) 54.04 200.00Avicel PH101 (Intra-granular) 18.00 66.60 Sodium Lauryl Sulfate 1.003.70 (Intra-granular) Plasdone K29/32 (Intra-solution) 3.00 11.10Pearlitol 200SD (Extra-granular) 17.95 66.40 Ac-Di-Sol (Extra-granular)5.00 18.50 Cab-O-Sil (Extra-granular) 0.50 1.85 Magnesium Stearate(Vegetable 0.50 1.85 Grade) (Extra-granular) Purified Water** Total:100.00 370.00 *Adusted for potency **Removed during processing, not infinal formulation.

Example

The stability of Form I and Form II of CEM-101 was evaluated undervarious conditions, as shown in the following Tables. In each case,there was no observed change in color of the test material during theentire study. In each case, the test material was first placed inPrimary Packaging Material (LLDPE bag within a HMHDPE/LDPE/LDPE blendbag, and then heat sealed). The Primary Packaging Material was placed-inSecondary Packaging Material (HDPE drum). All other impurities were lessthan 1% at each time point.

Form II, 2-8° C.

Initial 3 Month 6 Month 9 Month 12 Month Water 1.01% 1.24% 1.39% 1.38%1.37% Chromatographic Purity 97.0 96.0 96.8 96.4 96.8 3-EthynylanilineNot Not Not 0.01% 0.12% Detected Detected Detected Assay 93.9 93.4 93.693.3 93.0 (on hydrous basis)

Form II, 25° C./60% RH

Initial 3 Month 6 Month 9 Month 12 Month Water 1.01% 1.16% 1.42% 1.45%1.48% Chromatographic 97.0% 96.0% 96.7% 96.2% 96.1% Purity3-Ethynylaniline Not Not Not 0.04% 0.11% Detected Detected DetectedAssay 93.9% 93.6% 93.7% 93.2% 93.0% (on hydrous basis)

Form II, 40° C./75% RH

Initial 3 Month 6 Month 9 Month 12 Month Water 1.01% 1.24% 1.62% 1.51%1.56% Chromatographic Purity 97.0% 95.9% 96.5% 96.2% 97.1%3-Ethynylaniline Not Not Not 0.02% 0.05% Detected Detected DetectedAssay 93.9% 93.0% 93.1% 92.5% 92.3% (on hydrous basis)

Form I (Lot 3), 2-8° C.

Initial 3 Month 6 Month 12 Month Water 1.11% 1.63% 1.61% 1.69%Chromatographic 98.3% 98.1% 98.4% 98.5% Purity 3-Ethynylaniline 0.01%0.01% Not Detected Not Detected Assay 99.5% 98.9% 98.8% 98.7% (onhydrous basis)

Form I (Lot 3), 25° C./60% RH

Initial 3 Month 6 Month 12 Month Water 1.11% 1.65% 1.71% 1.75%Chromatographic 98.3% 98.1% 98.3% 98.4% Purity 3-Ethynylaniline 0.01%0.01% Not Detected Not Detected Assay 99.5% 98.7% 98.6% 98.5% (onhydrous basis)

Form I (Lot 3), 40° C./75% RH

Initial 3 Mouth 12 Month Water 1.11% 1.68% 1.84% Chromatographic 98.3%98.0% 98.3% Purity 3-Ethynylaniline 0.01% 0.01% 0.01% Assay 99.5% 98.5%98.3% (on hydrous basis)

Form I (Lot 4), 2-8° C.

Initial 3 Month 6 Month 12 Mouth Water 0.92% 1.56% 1.47% 1.51%Chromatographic 98.2% 98.1% 98.3% 98.4% Purity 3-Ethynylaniline 0.01%0.01% 0.01% Not Detected Assay 99.0% 98.4% 98.3% 98.2% (on hydrousbasis)

Form I (Lot 4), 25° C./60% RH

Initial 3 Month 6 Month 12 Month Water 0.92% 1.59% 1.75% 1.78%Chromatographic 98.2% 98.1% 98.4% 98.4% Purity 3-Ethynylaniline 0.01%0.01% Not Detected Not Detected Assay 99.0% 98.3% 98.2% 98.1% (onhydrous basis)

Form I (Lot 4), 40° C./75% RH

Initial 3 Month 12 Month Water 0.92% 1.65% 1.82% Chromatographic 98.2%98.4% 98.3% Purity 3-Ethynylaniline 0.01% 0.01% 0.01% Assay 99.0% 98.2%98.1% (on hydrous basis)

1.-32. (canceled)
 33. A composition comprising solithromycin incrystalline form characterized by differential scanning calorimetryhaving an endotherm at about 170° C., an endotherm in the range fromabout 194° C. to about 199° C., an endotherm in the range from about219° C. to about 225° C., an endotherm at about 225° C., or acombination thereof.
 34. The composition of claim 33 characterized bydifferential scanning calorimetry having an endotherm in the range fromabout 197° C. to about 198° C.
 35. The composition of claim 33characterized by differential scanning calorimetry having an endothermat about 225° C.
 36. The composition of claim 33 characterized bydifferential scanning calorimetry having an endotherm in the range fromabout 194° C. to about 199° C., and an endotherm in the range from about219° C. to about 225° C.
 37. A composition comprising solithromycin incrystalline Form I characterized by X-ray powder diffraction patternincluding a peak at ° 2θ=6.2, 8.5, or 8.8, each ±0.2 °2θ, or acombination thereof; and optionally solithromycin in crystalline Form IIcharacterized by X-ray powder diffraction pattern including a peak at °2θ=5.6 or 7.9, each ±0.2 °2θ, or a combination thereof.
 38. Thecomposition of claim 37 wherein the ratio of Form I to Form II is about90:10 or greater.
 39. The composition of claim 37 wherein the ratio ofForm I to Form II is about 95:5 or greater.
 40. The composition of claim37 wherein the ratio of Form I to Form II is about 98:2 or greater. 41.The composition of claim 37 wherein the ratio of Form I to Form II isabout 99:1 or greater.
 42. A composition comprising solithromycin incrystalline Form II characterized by X-ray powder diffraction patternincluding a peak at ° 2θ=5.6 or 7.9, each ±0.2 °2θ, or a combinationthereof; and optionally solithromycin in crystalline Form Icharacterized by X-ray powder diffraction pattern including a peak at °2θ=6.2, 8.5, or 8.8, each ±0.2 °2θ, or a combination thereof.
 43. Thecomposition of claim 42 wherein the ratio of Form II to Form I is 90:10or greater.
 44. The composition of claim 42 wherein the ratio of Form IIto Form I is 95:5 or greater.
 45. The composition of claim 42 whereinthe ratio of Form II to Form I is 98:2 or greater.
 46. The compositionof claim 42 wherein the ratio of Form II to Form I is 99:1 or greater.47. A method of treatment of a bacterial infection, a protozoalinfection, or a disorder caused at least in party by a bacterialinfection or protozoal infection in a host animal, the method comprisingthe step of administering to the host animal a therapeutically effectiveamount of the composition of claim 33, optionally further comprising atleast one pharmaceutically acceptable carrier, diluent, or excipient, ora combination thereof.